Tommaso Bonfiglio; Guendalina Olivero; Elisa Merega; Silvia Di Prisco; Cristina Padolecchia; Massimo Grilli; Marco Milanese; Lorenzo Di Cesare Mannelli; Carla Ghelardini; Giambattista Bonanno; Mario Marchi; Anna Pittaluga

“ Introduction Multiple sclerosis (MS) is mediated by an immune attack directed at myelin, which leads to a progressively degenerating disorder of the central nervous system (CNS) . Although immunological mechanisms are responsible for the majority of the cascade of events leading to MS, pathogenetic events involving neurons and astrocytes have been recently implicated in the pathogenesis of this disease [ [1] – [2] ] .
More precisely, recent work has targeted glutamate and GABA transmission at chemical synapsis in the CNS of EAE mice and MS patients. ”

“ In vivo prophylactic (0.3 mg/kg for 14 days, from the 7th day post immunization, d.p.i, the drug dissolved in the drinking water) fingolimod significantly reduced the clinical symptoms and the anxiety-related behaviour in EAE mice. Spinal cord inflammation, demyelination and glial cell activation are markers of EAE progression. These signs were ameliorated following oral fingolimod administration. Glutamate exocytosis was shown to be impaired in cortical and spinal cord terminals isolated from EAE mice at 21 ± 1 d.p.i., while GABA alteration emerged only at the spinal cord level. ”

“ Release studies Purified synaptosomes were prepared by homogenizing the cortex, the hippocampi and the spinal cord [ [26] ] of control and EAE mice in 10 volumes of 0.32 M sucrose, buffered to pH 7.4 with Tris- (hydroxymethyl) -amino methane [Tris, final concentration (f.c.) 0.01 M] [ [27] ] . The homogenate was centrifuged at 1,000 x g for 5 min, and the supernatant was stratified on a discontinuous Percoll gradient (2%, 6%, 10% and 20% v/v in Tris-buffered sucrose) and centrifuged at 33,500 x g for 5 min. ”